Fig. 3

Clec7a knockdown relieved neuropathic pain by regulating NLRP3 inflammasome-induced pyroptosis in vivo. A Timeline schematic of experimental paradigm. B Administration of Clec7a siRNA significantly ameliorated mechanical withdrawal threshold in CCI rat model (n = 10 rats/group). C The upregulation of Clec7a in spinal cord (SC) of CCI rats were significantly suppressed by si-Clec7a transfection on postoperative day 10 (n = 3 independent blots). D Immunohistochemical detection and quantification of Clec7a in ipsilateral SC of CCI rats after si-Clec7a treatment (n = 6 rats/group, 1 section/rat). White arrows point to Clec7a-positive cells. Scale bar: 100 μm (20 ×), 50 μm (40 ×). E Pyroptosis-related proteins (NLRP3, GSDMD) and pSyk, pERK, pJNK in ipsilateral SC of Sham or CCI rats treated with indicated siRNA were evaluated by western blotting (n = 3 independent blots). F The levels of IL-1β and IL-18 in serum of indicated groups were assessed by ELISA (n = 3 rats/group with 3 technical replicates). G Representative images (left) and quantification evaluation (right) of Tunel staining in indicated groups (n = 5 rats/group, 1 section/rat). Red: Tunel-positive cells; blue: nuclei (Hoechst 33,258). Scale bar: 100 μm. Data are presented as mean ± SEM. Two-way repeated measures ANOVA and Tukey’s post hoc test was used to analyze data at different time points B. One-way ANOVA was used to analyze data among multiple groups followed by Tukey’s post hoc test for equal variances or Dunnett T3 post hoc test for unequal variances C–G. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001 compared with the Sham group; ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 compared with Sham + si-NC group; ^△P < 0.05, ^△△P < 0.01, ^△△△P < 0.001 compared with CCI + si-NC group